59-05-032 Proceeding

50 Proceedings of the Princess Maha Chakri Sirindhorn Congress PURIFICATION AND CHARACTERIZATION OF BACTERIAL CELLULASE ACTIVITY OF BACILLUS FOR BIOFUEL PRODUCTION Passanee Thongthawee 1 , Kraipat Cheenkachorn 1 , Alisa Tolae 2 , Prapakorn Tantayotai 3 and Malinee Sriariyanun 4 Abstract : Lignocellulosic biomass is a renewable, abundant, and inexpensive resource for biorefining process. The aim of this study is to purify and characterize a potential thermotolerant bacterial cellulase to be further applied in biofuel production from lignocellulosic biomass. In our previous study, one thermotolerant bacterial strain, Bacillus sp. strain MSL2-S2 was screened based on cellulase assay from soil in the rice paddle field. Here, its cellulase enzyme was prepared by ammonium sulfate precipitation and fractionated by using size exclusion chromatography, sephacryl S-100 HR column. The molecular weight of purified cellulase was 50 kDa as revealed by SDS-PAGE and zymogram analysis. The optimal temperature and pH for enzyme activity was determined to be 55°C and pH 6.0. These properties of cellulase make it becomes potential to be applied in biomass conversion into fuel and other industrial processes. Keywords : Biofuel, Hydrolysis, Cellulase, Purification I. Introduction The increasing demand for energy in the industrial sector incurs global warming by use of fossil fuels, so the researchers tried to switch to the use of the alternative energy. Nowadays biofuel production from lignocellulosic biomass has gained more interest as a substitute for the use of fossil fuel. Lignocellulosic biomass is the most abundant renewable biomass in the world. It is the main composition of agricultural wastes, therefore by using lignocellulosic biomass as the material for biofuel production will have the additional benefit of helping to manage waste obtained from agricultural activities [1]. In general, the limiting step of biofuel production is the hydrolysis of lignocellulosic biomass because the cellulase enzyme has limited access to the biomass [2]. Cellulase is a complex enzyme system composing of endoglucanase, exoglucanase and β -glucosidase [3]. Bacterial cellulase is one of the most important options for enzyme production, because the bacteria have high growth 1 Department of Chemical Engineering, Faculty of Engineering, King Mongkut’s University of Technology, North Bangkok University 2 Department of Mechanical and Process Engineering, TGGS, King Mongkut’s University of Technology, North Bangkok University 3 Department of Biology, Srinakharinwirot University 4 Department of Mechanical and Process Engineering, TGGS, King Mongkut’s University of Technology, North Bangkok University.